System and method for forming synthetic protein crystals to determine the conformational structure by crystallography

Abstract

A method for forming synthetic crystals of proteins in a carrier fluid by use of the dipole moments of protein macromolecules that self-align in the Helmholtz layer adjacent to an electrode. The voltage gradients of such layers easily exceed 10 6 V/m. The synthetic protein crystals are subjected to x-ray crystallography to determine the conformational structure of the protein involved.

The invention claimed is: 
     
       1. A method for determining the conformational structure of a protein sample, the method comprising the steps of: applying an electrical voltage between two electrodes that interface with a liquid and protein macromolecule mixture;   maintaining said electrical voltage to promote formation of said protein macromolecules into pearl chains and synthesized three-dimensional protein crystals; and   screening said synthesized three-dimensional protein crystals by x-ray crystallography to determine the conformational structure of said basic protein.   
     
     
       2. The method of claim 1, wherein: the step of applying further comprises preventing an electrical current flow through said liquid and protein macromolecule mixture between said electrodes.   
     
     
       3. The method of claim 1, wherein: the step of applying further comprises a dielectric coating on said electrodes for preventing an electrical current flow through said liquid and protein macromolecule mixture between said electrodes.   
     
     
       4. The method of claim 1, wherein: the step of applying further comprises an orthogonal magnetic field for preventing an electrical current flow through said liquid and protein macromolecule mixture disposed between said electrodes.   
     
     
       5. The method of claim 1, wherein: the step of applying includes establishing an electric field strength between said electrodes in the range of three megavolts per meter to three gigavolts per meter and a parallel separation of said electrodes that does not exceed one micrometer.   
     
     
       6. The method of claim 1, wherein: the protein macromolecules are no smaller than one nanometer and no larger than one micrometer.   
     
     
       7. The method of claim 1, wherein: the step of applying further comprises a liquid and a protein macromolecule mixture that have substantially different dielectric constants.   
     
     
       8. The method of claim 1, wherein: the applying of the electric voltage is such that said mixture includes a substantial dielectric mismatch between the dielectric constant of the protein macromolecules and said liquid, and provides for the induction of a dipole moment.   
     
     
       9. A method for protein crystallography, the method comprising the steps of: diagnosing electric fields in a double layer of an electrode-fluid interface using electrochemistry techniques;   seeding this layer with polymer macromolecules and demonstrating that a complex fluid at the interface solidifies under the action of the electric field;   using electron microscopy to examine a registry of the macromolecules;   repeating the steps of seeding and using electron microscopy with a globular protein;   using x-ray scattering to see if the diffraction pattern of the globular proteins can be deconvolved to Angstrom resolution by computational modeling; and   comparing the resulting conformation with a preexisting protein database.   
     
     
       10. A method for creating diffraction-quality protein crystals on a microchip suitable for x-ray and electron diffraction studies, the method comprising the steps of: suspending a plurality of protein macromolecules each with a dipole moment in a liquid solution that in turn is disposed within a micron-sized gap between two micro-electrodes on a silicon substrate;   applying a voltage across said two micro-electrodes such that said protein macromolecules are aligned by the effects of an electric field in the electric double layer on permanent and induced dipole moments; and   electromechanical erecting at least one two-dimensional seed matrix providing for a subsequent self-assembly of at least one three-dimensional protein crystal.